SBIR-STTR Award

The objective is to develop new in situ (hybridization and immunocytochemistry) assay technology to take full advantage of the new generation of reagents for detecting low-abundance regulatory macromolecules in cells. In situ assays often defer to biochemical assays even though they are better suited, because they are less convenient and reliable. Standard in situ methods are capillarity-based, open-chamber methods characterized by handling and dehydration artifacts, poor temperature control, and many manual steps. Automated units that are now available work on the same principle and, consequently, do little more than compound the problems intrinsic to the capillarity design. The probe-clip reaction unit deviates from existing models in six key aspects:(1) the reaction chamber is closed rather than open,(2) there are two chambers rather than one,(3) in-chamber reactant mixing is by gravity rather than capillarity,(4) reactants are conveyed to the chamber in solid rather than liquid state,(5) washing is in a high-ambient-volume closed chamber rather than a fastflowing open stream, and(6) temperature control is by conduction rather than by convection.These changes give numerous advantages related to speed, safety, versatility, and sensitivity. In Phase I, a six-place manually operated prototype will be built for the research laboratory to validate the design concept for an automated model to be developed in Phase II.Awardee's statement of the potential commercial applications of the research:The manually operated unit will become standard equipment in most biology research laboratories. The automated unit will become the technology of choice in clinical laboratories. The device will make in situ assays easier in all applied sciences.National Institute of General Medical Sciences (NIGMS)

Our objective is to develop the scientific and engineering basis for a new reaction chamber technology that will be incorporated into a workstation for analytical cytochemistry. The technology is based on a "Probe-Clip" device, a disposable solid-phase reactant plate which seals reversibly to a tissue-bearing microscope slide to form a closed high-aspect(500 pm in height) reaction unit in which reactants flow by gravity. Preformatted lyophilized probe is sequestered in the unit during the blocking phase of assay and is activated on-command by rotating the closed unit in an Assembler to bring blocking solution into contact. The new technology addresses the three aspects of commercially available cytochemical assay technology which mitigate their use for quantitative work:(1) an open reaction chamber,(2) a low-aspect (ca. 150um) reaction chamber characterized by capillarity flow and large static boundary layers of reagents, and(3) ad hoc manipulation and delivery of probe.Phase I feasibility experiments showed that the process control afforded by Probe-Clip technology is better suited to the demands of analytical cytochemistry for low-abundance regulatory targets, using non-isotopic end-points. In Phase II, we will develop the scientific basis for reproducible fabrication of Probe-Clip and Assembler hardware, and to develop standard procedures for the use of the technology. Automation of the technology in Phase III will build on the culmination of Phase II research, a prototype robotic cassette mechanism.Awardee's statement of the potential commercial applications of the research:Our goal is to introduce a new reaction chamber technology to facilitate analytical cytochemistry and thereby to enlarge the market for in situ assays by increasing their accuracy for low-abundance regulatory targets (vis. receptors). Clinical pathology and research laboratories will benefit.National Institute of General Medical Sciences (NIGMS)